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pGLO Bacterial Transformation Kit - Bio-Rad

pGLO Bacterial Transformation Kit - Bio-Rad

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Published by ebujak
Biotechnology Explorer

Bio-Rad's pGLO(TM) Bacterial Transformation Kit

Biotechnology Explorer

Bio-Rad's pGLO(TM) Bacterial Transformation Kit

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Published by: ebujak on Apr 15, 2009
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05/11/2014

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Biotechnology Explorer
 ™ 
pGLO
 ™ 
BacterialTransformation Kit
Catalog Number166-0003EDUexplorer.bio-rad.com
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call
1-800-4BIORAD
(1-800-424-6723)
pGLO
 araCGFP blaori 
Store components of this kit at room temperature.
Duplication of any part of this document is permitted for classroom use only.
 
How can jellyfish shed light on the subject?
One of the biggest challenges for first-time students of biotechnology or molecular biology is thatmany of the events and processes they are studying are invisible. The Biotechnology Explorer  program has a solution: a gene from a bioluminescent jellyfish and its Green Fluorescent Protein—GFP.GFP fluoresces a brilliant green when viewed with a hand-held long-wave ultraviolet light (such as a pocket geology lamp).The gene for GFP was originally isolated from the jellyfish,
 Aequorea victoria
.The wild-type jellyfishgene has been modified by Maxygen Inc., a biotechnology company in Santa Clara, California.Specific mutations were introduced into the DNA sequence, which greatly enhance fluorescence of the protein. This modified form of the GFP gene has been inserted into Bio-Rad’s pGLO plasmidand is now available exclusively from Bio-Rad for educational applications.GFP is incredibly bright. Using pGLO to transform bacteria, students can actually observe geneexpression in real time. Following the transformation with Bio-Rad’s GFP purification kit, students purify the genetically engineered GFP from their transformed bacteria using a simple chromatography procedure. The entire process is visible using the hand-held UV lamp.
Guided Investigation
The intent of this curriculum is to guide students through the thought process involved in alaboratory-based scientific procedure. The focus here is not so much on the answer or result, butrather on how the result was obtained and how it can be substantiated by careful observation andanalysis of data. This is referred to as a guided inquiry-based laboratory investigation.At each step along the way, student understanding of the process and the analysis of data is stressed.Instead of providing students with explanations or interpretations, the Student Manual poses a seriesof questions to focus and stimulate thinking about all aspects of the investigation. Answers are providedin the Instructor’s Answer Guide.Student involvement in this process will result in an increased understanding of the scientific process and the value of proceeding into a task in an organized and logical fashion.Furthermore, we are expecting that students who engage in this type of process will start todevelop a more positive sense of their ability to understand the scientific method.Bio-Rad’s GFP-based curriculum is unique and has generated an unprecedented level of excitementamong science educators. We strive to continually improve our curriculum and products. Your inputis extremely important to us. We welcome your stories, comments, and suggestions.Ron MardigianDirector, Biotechnology Explorer program, Bio-Rad Laboratoriesron_mardigian@bio-rad.com
 
Table of Contents
Instructor’s Guide
Page
Introduction to Transformation............................................................................................1The pGLO System................................................................................................................1Kit InventoryChecklist........................................................................................................2Implementation Timeline......................................................................................................3Lesson Points to Highlight....................................................................................................3General Laboratory Skills......................................................................................3Experimental Points..............................................................................................5Conceptual Points..................................................................................................5Instructor’s Advance Preparation Overview........................................................................7Workstation Checklist..........................................................................................................7Instructor’s Advance Preparation Guide..............................................................................9Quick Guide (Graphic Laboratory Protocol)......................................................................14Instructor’s Answer Guide..................................................................................................16
Student Manual
Lesson 1Introduction to Transformation..................................................................28Focus Questions..........................................................................................29Lesson 2Transformation Laboratory........................................................................32Review Questions......................................................................................38Lesson 3Data Collection and Analysis....................................................................39Analysis of Results....................................................................................40Review Questions......................................................................................41Lesson 4ExtensionActivity: Calculate Transformation Efficiency........................43
Appendices
Appendix AHistorical Links to Biotechnology............................................................49Appendix BGlossary of Terms......................................................................................53Appendix CBasic Molecular Biology Concepts and Terminology..............................55Appendix DGene Regulation........................................................................................60Appendix EReferences..................................................................................................62

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